WashU weekly Neuroscience publications

CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
(2019) Scientific Reports, 9 (1), art. no. 9891, . 

Eaton, S.L.^a , Proudfoot, C.^a , Lillico, S.G.^a , Skehel, P.^{b c} , Kline, R.A.^a , Hamer, K.^e , Rzechorzek, N.M.^{d e} , Clutton, E.^f , Gregson, R.^f , King, T.^a , O’Neill, C.A.^g , Cooper, J.D.^{h i} , Thompson, G.^b , Whitelaw, C.B.^a , Wishart, T.M.^{a c}

^a The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, United Kingdom
^b Centre for Discovery Brain Science, University of Edinburgh, Hugh Robson Building, Edinburgh, United Kingdom
^c Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, United Kingdom
^d Centre for Clinical Brain Sciences, University of Edinburgh, Chancellor’s Building, 49 Little France Crescent, Edinburgh, United Kingdom
^e Royal (Dick) School of Veterinary Studies, The University of Edinburgh, Edinburgh, United Kingdom
^f Wellcome Trust Critical Care Laboratory for Large Animals, Roslin Institute, Easter Bush, Edinburgh, United Kingdom
^g BioMarin Pharmaceutical Inc, San Rafael, CA, United States
^h Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, David Geffen School of Medicine, UCLA, Torrance, CA, United States
ⁱ Department of Pediatrics, Washington University School of Medicine, St Louis, MO, United States

Abstract
The neuronal ceroid lipofuscinoses (NCLs) are a group of devastating monogenetic lysosomal disorders that affect children and young adults with no cure or effective treatment currently available. One of the more severe infantile forms of the disease (INCL or CLN1 disease) is due to mutations in the palmitoyl-protein thioesterase 1 (PPT1) gene and severely reduces the child’s lifespan to approximately 9 years of age. In order to better translate the human condition than is possible in mice, we sought to produce a large animal model employing CRISPR/Cas9 gene editing technology. Three PPT1 homozygote sheep were generated by insertion of a disease-causing PPT1 (R151X) human mutation into the orthologous sheep locus. This resulted in a morphological, anatomical and biochemical disease phenotype that closely resembles the human condition. The homozygous sheep were found to have significantly reduced PPT1 enzyme activity and accumulate autofluorescent storage material, as is observed in CLN1 patients. Clinical signs included pronounced behavioral deficits as well as motor deficits and complete loss of vision, with a reduced lifespan of 17 ± 1 months at a humanely defined terminal endpoint. Magnetic resonance imaging (MRI) confirmed a significant decrease in motor cortical volume as well as increased ventricular volume corresponding with observed brain atrophy and a profound reduction in brain mass of 30% at necropsy, similar to alterations observed in human patients. In summary, we have generated the first CRISPR/Cas9 gene edited NCL model. This novel sheep model of CLN1 disease develops biochemical, gross morphological and in vivo brain alterations confirming the efficacy of the targeted modification and potential relevance to the human condition. © 2019, The Author(s).

Document Type: Article
Publication Stage: Final
Source: Scopus
Access Type: Open Access

Optic Nerve Head Drusen: The Relationship Between Intraocular Pressure and Optic Nerve Structure and Function
(2018) Journal of neuro-ophthalmology : the Official Journal of the North American Neuro-Ophthalmology Society, 38 (2), pp. 147-150. Cited 2 times.

Nolan, K.W., Lee, M.S., Jalalizadeh, R.A., Firl, K.C., Van Stavern, G.P., McClelland, C.M.

Department of Ophthalmology and Visual Neurosciences (KWN, MSL, KCF, CMM), University of Minnesota School of Medicine, Minneapolis, Minnesota; and Department of Ophthalmology and Visual Sciences (RAJ, GPVS), Washington University School of Medicine, St. Louis, Missouri

Abstract
OBJECTIVE: To determine whether at the time of diagnosis, the intraocular pressure (IOP) in patients with optic nerve head drusen (ONHD) correlates with the perimetric mean deviation (PMD) and the mean retinal nerve fiber layer (RNFL) thickness on optical coherence tomography (OCT). METHODS: This retrospective chart review included adults with ONHD from 2 academic medical centers. Inclusion criteria were age older than 18 years, definitive diagnosis of ONHD, measurement of IOP, and an automated visual field (VF) within 3 months of diagnosis. Exclusion criteria were unreliable VFs, use of IOP-lowering therapy, and visually significant ocular comorbidities. Data were collected from the initial visit. Age, IOP, method of diagnosis of ONHD, mean RNFL thickness, and PMD were recorded. Multiple and logistic regression models were used to control for potential confounders in statistical analyses. RESULTS: Chart review identified 623 patients, of which 146 patients met inclusion criteria. Mean age was 44.2 years (range: 19-82 years). Average PMD of 236 eyes was -5.22 dB (range, -31.2 to +1.21 dB). Mean IOP was 15.7 mm Hg (range: 6-24 mm Hg). Forty eyes (16.9%) underwent RNFL measurement using OCT; mean RNFL thickness was 79.9 μm (range: 43-117 μm). There was no statistically significant association between IOP and PMD (P = 0.13) or RNFL thickness (P = 0.65). Eyes with ocular hypertension tended to have less depressed PMD than those without (P= 0.031). Stratified analyses of visible and buried subgroups yielded similar results. CONCLUSIONS: Lowering IOP in patients with ONHD has been proposed as a means to prevent progression of optic neuropathy. Our study demonstrated that among predominately normotensive eyes, higher IOP was not associated with greater VF loss or thinner RNFL at the time of presentation. This suggests that lowering IOP may not be beneficial in preventing visual loss in normotensive eyes with ONHD.

Document Type: Article
Publication Stage: Final
Source: Scopus